Efficient generation of bispecific IgG antibodies by split intein mediated protein trans-splicing system

Abstract Many methods have been developed to produce bispecific antibodies (BsAbs) for industrial application. However, huge challenges still remain in synthesizing whole length BsAbs, including their assembly, stability, immunogenicity, and pharmacodynamics. Here we present for first time a generic...

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Autores principales:	Lei Han, Junsheng Chen, Kai Ding, Huifang Zong, Yueqing Xie, Hua Jiang, Baohong Zhang, Huili Lu, Weihan Yin, John Gilly, Jianwei Zhu
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/94d07bcb19cd4ac6bb1c314751914989
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spelling	oai:doaj.org-article:94d07bcb19cd4ac6bb1c3147519149892021-12-02T15:05:09ZEfficient generation of bispecific IgG antibodies by split intein mediated protein trans-splicing system10.1038/s41598-017-08641-32045-2322https://doaj.org/article/94d07bcb19cd4ac6bb1c3147519149892017-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-08641-3https://doaj.org/toc/2045-2322Abstract Many methods have been developed to produce bispecific antibodies (BsAbs) for industrial application. However, huge challenges still remain in synthesizing whole length BsAbs, including their assembly, stability, immunogenicity, and pharmacodynamics. Here we present for first time a generic technology platform of generating bispecific IgG antibodies, “Bispecific Antibody by Protein Trans-splicing (BAPTS)”. Different from published methods, we assembled two parental antibody fragments in the hinge region by the protein trans-splicing reaction of a split intein to generate BsAbs without heavy/heavy and light/heavy chain mispairing. Utilizing this simple and efficient approach, there have been several BsAbs (CD3×HER2, CD3×EGFR, EGFR×HER2) synthesized to demonstrate its broad applicability. Correctly paired mAb arms were assembled to form BsAbs that were purified through protein A affinity chromatography to demonstrate industrial applicability at large scale. Further, the products were characterized through physical-biochemistry properties and biological activities to confirm expected quality of the products from “BAPTS”. More importantly, correct pairing was confirmed by mass spectrum. Proof-of-concept studies with CD3×HER2 BsAb (T-cell recruitment) demonstrated superior bioactivity compared with trastuzumab. The results of undetectable mispairing and high biological activity have indicated that this method has the potential to be utilized to manufacture BsAbs with high efficiency at industrial scale.Lei HanJunsheng ChenKai DingHuifang ZongYueqing XieHua JiangBaohong ZhangHuili LuWeihan YinJohn GillyJianwei ZhuNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Lei Han Junsheng Chen Kai Ding Huifang Zong Yueqing Xie Hua Jiang Baohong Zhang Huili Lu Weihan Yin John Gilly Jianwei Zhu Efficient generation of bispecific IgG antibodies by split intein mediated protein trans-splicing system
description	Abstract Many methods have been developed to produce bispecific antibodies (BsAbs) for industrial application. However, huge challenges still remain in synthesizing whole length BsAbs, including their assembly, stability, immunogenicity, and pharmacodynamics. Here we present for first time a generic technology platform of generating bispecific IgG antibodies, “Bispecific Antibody by Protein Trans-splicing (BAPTS)”. Different from published methods, we assembled two parental antibody fragments in the hinge region by the protein trans-splicing reaction of a split intein to generate BsAbs without heavy/heavy and light/heavy chain mispairing. Utilizing this simple and efficient approach, there have been several BsAbs (CD3×HER2, CD3×EGFR, EGFR×HER2) synthesized to demonstrate its broad applicability. Correctly paired mAb arms were assembled to form BsAbs that were purified through protein A affinity chromatography to demonstrate industrial applicability at large scale. Further, the products were characterized through physical-biochemistry properties and biological activities to confirm expected quality of the products from “BAPTS”. More importantly, correct pairing was confirmed by mass spectrum. Proof-of-concept studies with CD3×HER2 BsAb (T-cell recruitment) demonstrated superior bioactivity compared with trastuzumab. The results of undetectable mispairing and high biological activity have indicated that this method has the potential to be utilized to manufacture BsAbs with high efficiency at industrial scale.
format	article
author	Lei Han Junsheng Chen Kai Ding Huifang Zong Yueqing Xie Hua Jiang Baohong Zhang Huili Lu Weihan Yin John Gilly Jianwei Zhu
author_facet	Lei Han Junsheng Chen Kai Ding Huifang Zong Yueqing Xie Hua Jiang Baohong Zhang Huili Lu Weihan Yin John Gilly Jianwei Zhu
author_sort	Lei Han
title	Efficient generation of bispecific IgG antibodies by split intein mediated protein trans-splicing system
title_short	Efficient generation of bispecific IgG antibodies by split intein mediated protein trans-splicing system
title_full	Efficient generation of bispecific IgG antibodies by split intein mediated protein trans-splicing system
title_fullStr	Efficient generation of bispecific IgG antibodies by split intein mediated protein trans-splicing system
title_full_unstemmed	Efficient generation of bispecific IgG antibodies by split intein mediated protein trans-splicing system
title_sort	efficient generation of bispecific igg antibodies by split intein mediated protein trans-splicing system
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/94d07bcb19cd4ac6bb1c314751914989
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_version_	1718388907153817600

Efficient generation of bispecific IgG antibodies by split intein mediated protein trans-splicing system

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